RNA干扰β-catenin基因对二氧化硅诱导的MH-S细胞中基质金属蛋白酶表达的影响

Recombinant lentivirus containing β-catenin short hairpin RNA inhibits expressionin of MMP in MH-S cells after silica treatment

目的观察β-catenin短发夹RNA干扰(shRNA)重组慢病毒对二氧化硅诱导的小鼠肺泡巨噬细胞系MH-S细胞中基质金属蛋白酶(Matrix Metalloproteinases,MMP)9表达的影响。方法将MH-S细胞分为4组:空白细胞对照组(MH-S)、SiO_2诱导组(silica)、病毒干扰组(silica+LV-shβ-catenin)、病毒对照组(silica+LV-shβ-catenin-NC),除空白细胞对照组外,各组细胞均用终浓度为200μg/ml的SiO_2刺激,培养18 h后,western blot检测各组细胞β-catenin和MMP9蛋白表达水平,realtime-PCR检测各组细胞MMP9 mRNA水平。结果 SiO_2诱导后,与silica组和silica+LV-shβ-cateninNC组比较,silica+LV-shβ-catenin组β-catenin蛋白表达显著降低(P<0.05);realtime-PCR和western blot检测结果显示,silica+LV-shβ-catenin组细胞中MMP9的mRNA和蛋白表达较silica组和silica+LV-shβ-catenin-NC组均显著降低(P<0.05)。结论重组慢病毒LV-shβ-catenin能够有效抑制经SiO_2诱导的MH-S细胞株β-catenin蛋白表达,并对MMP9的表达有明显抑制作用。

Objective To study the inhibitory effect ofβ-catenin targeted RNA interference on matrix metalloproteinases expression in MH-S cells,mouse alveolar macrophages,induced by silica dust.Methods The MH-S cells were divided into four groups：the MH-S group,silica group,silica ＋LV-shβ-catenin（ a lentiviral vector expressing shRNA specific against mouse β-catenin） group and silica ＋LV-shβ-catenin-NC（ non-silence control lentivirus） group.Each group of cells was induced by silica dust at the final concentration of 200 ug/ml and incubated for 18 h.The expressions of β-catenin and MMP9 were detected by Western blot and real-time PCR respectively.Results After induction by silica dust,the expression of β-catenin in the silica ＋LV-shβ-catenin group was significantly lower than that in the silica and silica ＋LV-shβ-catenin-NC group（ P〈0.05）.According to the result of Western blot and real-time PCR,the expressions of MMP9 in the silica ＋LV-shβ-catenin group was significantly lower than those in the silica and silica ＋LV-shβ-catenin-NC group（ P〈0.05）.Conclusion Lv-shβ-catenin can significantly inhibit β-catenin and MMP9 expression in MH-S cells activated by silica dust.